CN115120634A

CN115120634A - Traditional Chinese medicine compound preparation for preventing and treating alcoholic liver injury and preparation method and application thereof

Info

Publication number: CN115120634A
Application number: CN202110315877.XA
Authority: CN
Inventors: 谭昌恒; 曲世津; 高明垚; 李牧; 谭俊杰; 骆宏丰
Original assignee: Shanghai Institute of Materia Medica of CAS
Current assignee: Shanghai Institute of Materia Medica of CAS
Priority date: 2021-03-24
Filing date: 2021-03-24
Publication date: 2022-09-30
Anticipated expiration: 2041-03-24
Also published as: CN115120634B

Abstract

The invention relates to the technical field of medicaments and health-care foods, and particularly provides a composition which comprises an active ingredient A, B, C; wherein the active ingredient A is selected from the following group: dried root, stem, whole plant or extract thereof of Salvia miltiorrhiza; the active ingredient B is selected from the following group: dried flowers of safflower, or extracts thereof; the active ingredient C is selected from the group consisting of: vitamin C, vitamin E, or a combination thereof. When the active component C consists of vitamin C and vitamin E, the composition comprises the following components in parts by weight based on the total weight of the composition as 1000 parts: 350-600 parts of active ingredient A, 250-500 parts of active ingredient B, 45-60 parts of vitamin C and 8-18 parts of vitamin E. The composition provided by the invention is used for preventing and/or treating and/or slowing down alcoholic liver injury.

Description

Traditional Chinese medicine compound preparation for preventing and treating alcoholic liver injury and preparation method and application thereof

Technical Field

The application relates to the technical field of medicines and health-care foods, in particular to a traditional Chinese medicine compound preparation or health-care food for preventing and/or treating alcoholic liver injury and a preparation method and application thereof.

Background

Alcoholic Liver Disease (ALD) is toxic liver injury caused by long-term excessive drinking. When the fat is accumulated in liver cells for a long time and exceeds 5% of the wet weight of the liver, or when the liver cells with the unit area of 1/3 or more in histology have steatosis, the further development can cause alcoholic hepatitis and hepatic fibrosis, and finally, the liver cirrhosis is caused. Extensive hepatocellular necrosis and even liver failure are induced during severe alcohol abuse. ALD has become a common disease that seriously harms human health.

Modern medicine's treatment of alcoholic liver injury focuses mainly on alcohol withdrawal and nutritional support, reducing the severity of alcoholic liver injury and treatment for various complications. The only option for patients with advanced liver cirrhosis is liver transplantation, but due to various factors, there are few patients who have had the opportunity to receive liver transplantation therapy. To date, the FDA in the united states has not approved any therapeutic drug for alcoholic liver disease.

According to the basic theory of traditional Chinese medicine, the liver and spleen are the disease sites of alcoholic liver diseases, dampness, phlegm, qi stagnation and stasis are the main disease properties, and the commonly used treatment methods are heat clearing and dampness removing, liver soothing and qi regulating, blood circulation promoting and blood stasis removing, qi tonifying and spleen strengthening. At present, the traditional Chinese medicine has a plurality of researches for preventing and treating alcoholic liver diseases. However, the traditional Chinese medicine has slow curative effect, needs more dosage or is taken for a long time, and easily generates certain toxic and side effects on the damaged liver.

Therefore, the development of a high-safety Chinese herbal compound preparation for treating and/or relieving alcoholic liver diseases is urgently needed in the field.

Disclosure of Invention

The invention aims to develop a high-safety Chinese herbal compound preparation for treating and/or relieving alcoholic liver diseases. Specifically, the invention provides a traditional Chinese medicine compound preparation which mainly comprises salvia miltiorrhiza and safflower and is assisted by vitamin C and vitamin E, and is used for preventing and/or treating and/or relieving alcoholic liver injury. In addition, the invention also provides a preparation method of the traditional Chinese medicine compound preparation.

In a first aspect of the invention, there is provided a composition comprising an active ingredient A, B, C;

wherein the active ingredient A is selected from the following group: dried root, stem, whole plant or extract thereof of Saviae Miltiorrhizae radix;

the active ingredient B is selected from the following group: dried flowers of safflower, or extracts thereof;

the active ingredient C is selected from the following group: vitamin C, vitamin E, or a combination thereof.

In another preferred embodiment, the composition is a traditional Chinese medicine composition, and the active ingredient A and the active ingredient B are traditional Chinese medicinal materials.

In another preferred embodiment, when the active ingredient is an extract, the compatibility of the extract is calculated according to the dry weight of the corresponding medicinal materials.

In another preferred embodiment, the medicinal material comprises fresh Chinese medicinal materials, partially dried Chinese medicinal materials, dried Chinese medicinal materials or a combination thereof.

In another preferred example, when the active ingredients are traditional Chinese medicinal materials, the weight parts of the active ingredients are calculated according to the dry weight of the corresponding traditional Chinese medicinal materials.

In another preferred embodiment, when the active ingredient is an extract, the weight parts thereof are calculated according to the dry weight of the corresponding Chinese medicinal material.

In another preferred embodiment, the extract is an extract of water or an aqueous solvent.

In another preferred embodiment, when the active ingredient C consists of vitamin C and vitamin E, the composition comprises, by weight, 1000 parts of the total weight of the composition:

350-600 parts of active ingredient A, 250-500 parts of active ingredient B, 45-60 parts of vitamin C and 8-18 parts of vitamin E.

In another preferred embodiment, the "by weight" is based on dry weight.

In another preferred embodiment, the composition is an oral formulation.

In another preferred embodiment, the composition is a decoction of the Chinese medicinal materials.

In another preferred embodiment, the composition is a decoction.

In another preferred embodiment, the decoction comprises decoction with unadjusted (or unchanged) concentration, concentrated decoction and diluted decoction.

In a second aspect of the invention, there is provided a pharmaceutical formulation comprising i) a composition according to the first aspect;

ii) other pharmaceutically acceptable adjuvants, carriers or excipients.

In a third aspect of the invention, there is provided a use of the composition of the first aspect or the pharmaceutical preparation of the second aspect for preparing a medicament and/or health food for preventing and/or treating alcoholic liver disease.

In another preferred example, the alcoholic liver disease comprises alcoholic liver injury.

In another preferred embodiment, the pharmaceutical preparation is used for preparing a medicament with the following effects:

(X1) reducing the level of Triglycerides (TG) in serum;

(X2) reducing hepatic Malondialdehyde (MDA) content in liver tissue;

(X3) increased reduced Glutathione (GSH) levels in liver tissue.

In another preferred embodiment, the dosage form of the pharmaceutical preparation is selected from the group consisting of: oral liquid, tablet, granule, pill, and capsule.

In another preferred embodiment, the pharmaceutical formulation is a capsule.

In another preferred embodiment, the pharmaceutical formulation is an oral formulation.

In another preferred embodiment, the pharmaceutical formulation is a liquid formulation or a lyophilized formulation.

In a fourth aspect of the present invention, there is provided a method of preparing the pharmaceutical formulation of the second aspect, comprising the steps of:

s1) providing a salvia miltiorrhiza extract, a safflower extract;

s2) mixing and stirring a certain amount of compatible salvia miltiorrhiza extract, safflower extract, vitamin C, vitamin E and medically acceptable auxiliary materials to obtain a first mixture;

s3) mixing the first mixture with ethanol as a binder according to a certain compatibility to obtain a second mixture;

s4) sieving the second mixture with a 10-mesh sieve, and drying the preparation at 80 ℃ to obtain the pharmaceutical preparation.

In another preferred example, the compatibility in step S2) is: the total amount of the first mixture is 1000 parts, the red sage root extract accounts for 350-600 parts, the safflower extract accounts for 25-50 parts, the vitamin C accounts for 45-60 parts, and the vitamin E accounts for 8-18 parts.

In another preferred example, the binder ethanol in step S3) is preferably 40% ethanol.

In another preferred example, the compatibility in step S3) is: the first mixture, by weight: the ratio of ethanol is 2-5: 4-6.

In another preferred example, the formulation in step S4) includes: tablet, granule, pill, and capsule.

In another preferred example, the providing of the salvia miltiorrhiza extract in the step S1) includes the detailed steps of:

D1) under the heating condition, extracting a salvia miltiorrhiza medicinal material by using water for 45-90 min to obtain an extracting solution;

D2) repeating the step D1) for 2-3 times, and mixing the extracting solutions obtained each time to obtain a first extracting solution;

D3) concentrating the first extracting solution under reduced pressure, and adding 95% alcohol to obtain a first concentrated solution;

D4) standing the first concentrated solution overnight, and concentrating supernatant to obtain a second concentrated solution;

D5) spray drying the second concentrated solution to obtain the Saviae Miltiorrhizae radix extract.

In another preferred example, the volume of the water in the step D1) is 6-8 times (V/W) of the weight of the salvia miltiorrhiza bunge.

In another preferred embodiment, the heating temperature in step D1) is preferably 90 ℃.

In another preferred example, the duration in step D1) is preferably 60 min.

In another preferred example, the relative density of the concentrated first extract in the step D3) is 1.05-1.15.

In another preferred example, the alcohol concentration in the first concentrated solution in the step D3) is 60%.

In another preferred embodiment, the preferred operating parameters for spray drying in step D5) are: the inlet temperature is 140-150 ℃, and the outlet temperature is 50-60 ℃.

In another preferred example, the providing of the safflower extract in the step S1) includes the detailed steps of:

H1) under the heating condition, extracting a safflower medicinal material by using water for 30-75 min to obtain an extracting solution;

H2) repeating the step H1) 2-3, and mixing the extracting solutions obtained each time to obtain a second extracting solution;

H3) concentrating the second extractive solution under reduced pressure, and adding 95% ethanol to obtain third concentrated solution;

H4) standing the third concentrated solution overnight, and concentrating the supernatant to obtain a fourth concentrated solution;

H5) spray drying the fourth concentrated solution to obtain the safflower extract.

In another preferred example, the volume of the water in the step H1) is 10-15 times (V/W) of the weight of the safflower medicinal material.

In another preferred embodiment, the heating temperature in step H1) is preferably 80 ℃.

In another preferred example, the duration in step H1) is preferably 40 min.

In another preferred example, the relative density of the concentrated first extract in the step H3) is 1.05 to 1.15.

In another preferred example, the alcohol concentration in the first concentrated solution in the step H3) is 60%.

In another preferred example, the preferred operating parameters for spray drying in step H5) are: the inlet temperature is 115-125 ℃, and the outlet temperature is 50-60 ℃.

In a fifth aspect of the invention, there is provided a method of treating alcoholic liver disease, comprising the steps of: administering to a subject in need thereof a medically effective amount of the composition of the first aspect or the pharmaceutical formulation of the second aspect.

In another preferred example, the subject is an alcoholic liver patient.

In a sixth aspect of the invention, there is provided a method of treating alcoholic liver disease, comprising the steps of: administering to a subject in need thereof a medically effective amount of the composition of the first aspect or the pharmaceutical formulation of the second aspect.

In another preferred example, the subject is an alcoholic liver patient.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be repeated herein, depending on the space.

Detailed Description

The present inventors have made extensive and intensive studies and, as a result, have unexpectedly found that red sage root and safflower have a good effect in the treatment of alcoholic liver disease, and have completed the present invention.

The compound preparation takes 1000 parts as a base number, and comprises 350-600 parts of salvia extract, 250-500 parts of safflower extract, 45-60 parts of vitamin C, 8-18 parts of vitamin E and a proper amount of pharmaceutic adjuvant.

Term(s) for

As used herein, the terms "compound preparation of Chinese medicinal herbs", "formulation of the present invention", "preparation of the present invention", and "pharmaceutical preparation of the present invention" are used interchangeably and refer to a compound preparation of Chinese medicinal herbs based on Danshen root and safflower, supplemented with vitamin C and vitamin E, wherein the compound preparation contains, based on 1000 parts by weight, 350-600 parts by weight of Danshen root extract, 250-500 parts by weight of safflower extract, 45-60 parts by weight of vitamin C, 8-18 parts by weight of vitamin E, and a suitable amount of pharmaceutical excipients.

Alcoholic Liver Disease (ALD)

Modern medicine believes that ALD involves 4 basic pathological manifestations, (1) steatosis in large-vesicular or mixed hepatocytes, the earliest and most common occurrence; (2) hepatocellular injury, often manifested as balloon-like degeneration; (3) infiltration of liver lobule inflammation; (4) liver fibrosis and lobular architecture destruction to varying degrees.

ALD is mainly the result of the interaction of various factors such as inflammatory reaction, oxidative stress, enterogenic endotoxins, inflammatory mediators and nutrient imbalance, which are directly or indirectly induced in the metabolic process of ethanol and derivatives thereof. According to the "secondary stroke" theory, alcohol factors induce liver fat accumulation as a primary stroke by increasing reactive oxides through oxidative stress. Under the action of lipid peroxidation and inflammatory cytokines related to oxidative stress, fatty liver cells are subjected to secondary hit, so that inflammation, necrosis and fibrosis are caused.

Red sage root-safflower

The red sage-safflower is a famous blood-activating medicine pair, and the two medicines are both essential blood-activating medicines and enter heart and liver channels, and are used for removing blood stasis and promoting tissue regeneration and nourishing liver and kidney. Salvia miltiorrhiza, which is the dried root and rhizome of Salvia miltiorrhiza Bge (Salvia miliiorrhiza Bge.) belonging to Labiatae, was listed as the top grade in Shennong Ben Cao Jing, and is bitter in taste and slightly cold in nature, and enters heart and liver meridians. Has the effects of activating blood circulation to dissipate blood stasis, stimulating the menstrual flow to relieve pain, clearing away the heart-fire and relieving restlessness, cooling blood and eliminating carbuncle. Safflower is the dried flower of Carthamus tinctorius L of Compositae, is pungent and warm in taste, and is one of the main traditional Chinese medicines for promoting blood circulation and removing blood stasis. Pharmacological research shows that water-soluble phenolic acid components in the salvia miltiorrhiza, such as danshensu, rosmarinic acid, lithospermic acid, salvianolic acid B and the like, are the main effective components of the salvia miltiorrhiza.

Pharmaceutical formulations and methods of administration

The present invention also provides a pharmaceutical formulation comprising i) sexual ingredient A, B, C;

wherein the active ingredient A is selected from the following group: dried root, stem, whole plant or extract thereof of Salvia miltiorrhiza;

the active ingredient C is selected from the following group: vitamin C, vitamin E, or a combination thereof;

ii) other pharmaceutically acceptable adjuvants, carriers or excipients.

In another preferred embodiment, when the active ingredient C consists of vitamin C and vitamin E, the total weight of the composition is 1000 parts by weight, and the composition comprises:

The red sage root and safflower extracts are both extracts extracted and refined by safe media (water and alcohol);

the vitamin C and the vitamin E are nutrient supplements, fully embody the safety of the traditional Chinese medicine compound preparation, and are suitable for long-term administration.

As used herein, the term "effective amount" or "effective dose" refers to an amount that produces a function or activity (i.e., anti-aging function) in a human and/or animal and is acceptable to the human and/or animal.

As used herein, an ingredient of the term "pharmaceutically acceptable" is one that is suitable for use in humans and/or mammals without undue adverse side effects (such as toxicity, irritation, and allergic response), i.e., at a reasonable benefit/risk ratio. The term "pharmaceutically acceptable carrier" refers to a carrier for administration of a therapeutic agent, including various excipients and diluents.

The pharmaceutical composition of the present invention contains a safe and effective amount of the active ingredient of the present invention and a pharmaceutically acceptable carrier. Such vectors include (but are not limited to): saline, buffer, glucose, water, glycerol, ethanol, and combinations thereof. The dosage form of the pharmaceutical composition of the invention is injection, oral preparation (tablet, capsule, oral liquid), transdermal agent, sustained release agent. For example, by a conventional method using physiological saline or an aqueous solution containing glucose and other adjuvants. The pharmaceutical composition is preferably manufactured under sterile conditions.

The effective amount of the active ingredient of the present invention may vary depending on the mode of administration and the severity of the disease to be treated, etc. The selection of a preferred effective amount can be determined by one of ordinary skill in the art based on a variety of factors (e.g., by clinical trials). Such factors include, but are not limited to: pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated by the patient, the weight of the patient, the immune status of the patient, the route of administration, and the like. In general, satisfactory results are obtained when the active ingredient of the invention is administered at a daily dose of about 0.00001mg to 50mg per kg of animal body weight (preferably 0.0001mg to 10mg per kg of animal body weight). For example, divided doses may be administered several times per day, or the dose may be proportionally reduced, as may be required by the urgency of the condition being treated.

Typically, when the formulation of the invention is administered orally, 0mg, more preferably 50-250mg, is administered. The daily dose may be administered in one, two or more divided doses.

The pharmaceutically acceptable carrier of the present invention includes (but is not limited to): water, saline, liposomes, lipids, peptidic substances, cellulose, nanogels, or combinations thereof. The choice of carrier should be matched with the mode of administration, and these are well known to those skilled in the art.

Preparation method

The invention also provides a preparation method of the preparation, which comprises the following steps:

s1) providing a salvia miltiorrhiza extract, a safflower extract;

D3) concentrating the first extract under reduced pressure, and adding 95% ethanol to obtain a first concentrated solution;

In another preferred example, the duration in step D1) is preferably 60 min.

In another preferred example, the relative density of the concentrated first extract in the step D3) is 1.05-1.15

H3) concentrating the second extract under reduced pressure, and adding 95% ethanol to obtain a third concentrated solution;

In another preferred example, the heating temperature in step H1) is preferably 80 ℃.

In another preferred example, the duration in step H1) is preferably 40 min.

In another preferred example, the relative density of the concentrated first extract in step H3) is 1.05 to 1.15, and in another preferred example, the alcohol concentration in the concentrated first extract in step H3) is 60%.

The main advantages of the present invention include:

(1) the traditional Chinese medicine compound preparation is a preventive medicine-food homologous compound preparation suitable for daily administration, and has the characteristic of high safety while treating alcoholic liver diseases.

(2) The traditional Chinese medicine compound preparation disclosed by the invention can be used for effectively treating alcoholic liver diseases by reducing the content of MDA and TG in liver tissues and increasing the content of GSH.

(3) The invention creatively provides a traditional Chinese medicine compound preparation of salvia miltiorrhiza and safflower extract with a certain proportion and vitamin C and vitamin E with a certain proportion, and the compound preparation can effectively prevent and/or treat and/or alleviate alcoholic liver diseases.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, generally followed by conventional conditions, such as Sambrook et al, molecular cloning: the conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press,1989), or according to the manufacturer's recommendations. Unless otherwise indicated, percentages and parts are percentages and parts by weight.

Example 1 preparation of a Compound Chinese medicinal preparation for preventing alcoholic liver disease

1) Preparing a salvia miltiorrhiza extract: slicing and crushing 10kg of salvia miltiorrhiza, adding 8 times of water for the first time, soaking overnight, performing heat extraction at 90 ℃ for 1h, and sieving with a 80-mesh sieve to obtain a filtrate. Extracting twice, combining the filtrates, concentrating to a small volume, adding 95% alcohol until the alcohol concentration of the solution is 60%, standing overnight, collecting the supernatant, concentrating until the specific gravity is 1.05-1.15, and spray drying (inlet temperature is 140-150 ℃, outlet temperature is 50-60 ℃) to obtain the salvia miltiorrhiza extract.

2) Preparation of safflower extract: 10kg of safflower dried flower is added with 14 times of water, soaked overnight and heated to 80 ℃ for extraction for 40min, and then the safflower is sieved by a 80-mesh sieve to obtain filtrate. Repeating the extraction twice, combining the filtrates, concentrating to a small volume, adding 95% alcohol until the alcohol concentration of the solution is 80%, standing overnight, taking the supernatant, concentrating until the specific gravity is 1.05-1.15, concentrating, and performing spray drying (the inlet temperature is 115-125 ℃, and the outlet temperature is 50-60 ℃) to obtain the safflower extract.

3) Preparing a compound preparation mixed material: the red sage root extract, the safflower extract, the vitamin C, the vitamin E and the auxiliary material filler microcrystalline cellulose are sieved (100 meshes), mixed according to the proportion of 47.4 percent, 39.2 percent, 5.4 percent, 1.5 percent and 6.1 percent, and then put into a V-shaped mixer to be mixed for 30min to ensure the uniformity. Adding wetting agent 40% ethanol into the above mixed raw materials, making into soft mass, drying in boiling drier at 80 deg.C for 5min, grading with 100 mesh sieve, adding 0.5% lubricant magnesium stearate (sieving with 100 mesh sieve), and mixing to obtain granule.

4) Preparing capsules: the capsules were filled using a capsule filling machine with about 0.4g of capsule contents per capsule. Removing the fine powder adhered on the surface of the capsule by a polishing machine, and screening to remove unqualified products.

Example 2 construction of an animal model for alcoholic liver injury

Experimental ICR mice (female, 20 ± 2g in weight) were acclimatized for 4 days in the environment prior to the experiment. Experiments were subsequently performed. The model group mice are given deionized water and weighed once a week, and on the 30 th day of the experiment, the model group mice are given 50% ethanol 14mL/kg body weight and are subjected to intragastric administration once, so that a liver injury animal model is established.

Example 3 Effect of Compound Chinese medicinal preparation on animal model

3.1 Experimental methods: the compound preparation prepared in example 1 was used as an example for the study

(1) Experiment grouping

The test is provided with an alcoholic liver injury mouse model control group, a traditional Chinese medicine treatment group and a blank control group. The traditional Chinese medicine treatment groups are divided into a low-dose group (L), a medium-dose group (M) and a high-dose group (H), the doses of the groups are respectively (0.065, 0.13 and 0.4g/kg of body weight, which are respectively equal to 5, 10 and 30 times of recommended doses of people), the groups are orally administered by intragastric administration every day, and a blank control group and a model control group are administered with the same dose of deionized water.

(2) Correlation index detection

On the 30 th day of the experiment, the animals of the model control group and the test substance in each dose group were subjected to intragastric administration with 14mL/kg of 50% ethanol once, the animals of the blank control group were subjected to distilled water, the animals were sacrificed after fasting for 16 hours, and the liver lobes were taken for histopathological examination. Freezing the rest liver tissues with liquid nitrogen, and storing in a refrigerator at-40 deg.C for detecting various indexes.

Malondialdehyde (MDA), reduced Glutathione (GSH) and Triglycerides (TG) in liver tissue: adding 9 parts of cold normal saline into 1 part of liver tissue, homogenizing, centrifuging at 3000 r/min for 10 min, and taking supernatant for later use. The contents of MDA and GSH in liver tissue are determined by kit (provided by Nanjing institute of bioengineering), and TG content is analyzed by TBA-40FR full-automatic biochemical analyzer.

Histopathological examination of the liver: taking the left leaf of mouse liver as cross section, freezing and slicing, and staining Sudan III. The pathological changes of cells are recorded from one end of the liver by microscopic examination, and the whole tissue section is continuously observed by a 40-time objective lens, so that the distribution, the range and the area of lipid drops in the liver are mainly observed.

(3) Data processing

The experimental results were counted using SPSSll.5. The raw data were statistically analyzed using analysis of variance or t-test.

3.2 results of the experiment

(1) Effect of the compound preparation prepared in example 1 on body weight of experimental mice: as shown in table 1. The mice in the low, medium and high three-dose groups have no significant difference in initial weight, intermediate-stage weight, final-stage weight and weight gain compared with the model control group (analysis of variance, p > 0.05).

(2) The effect of the compound preparation prepared in example 1 on the contents of MDA, GSH and TG in liver tissues of mice: as shown in table 2. Compared with a blank control group, the liver tissues MDA and TG of the mice in the model control group are obviously increased, the GSH is obviously reduced, and the differences are all significant (t test, p is less than 0.05), which indicates that the alcoholic liver injury model is established, and compared with the model control group, the liver tissues MDA and TG contents of the mice in the medium and high dose groups are obviously reduced, the GSH of the mice in the medium dose group is obviously increased, and the differences are all significant (q test, p is less than 0.05).

(3) Effect of the compound preparation prepared in example 1 on the histopathological changes of the liver: as shown in table 3. The pathological change of the liver of the animals in the model control group and each dosage group is mainly liver cell steatosis, and the pathological change part is seen in the peripheral zone of lobules. Compared with a blank control group, the average value of the fatty degeneration score of the liver cells of the mouse in the model control group is obviously improved, and the difference is significant (t test, p is less than 0.05), which indicates that the model of the alcoholic liver injury is established. The average values of the fatty degeneration of liver cells of the mice in the four experimental groups have significance (analysis of variance, p is less than 0.05), but the average values of the fatty degeneration of liver cells of the mice in the three dose groups have no significance when being compared with the average value of the fatty degeneration of liver cells of the model control group (q test, p is more than 0.05). The test object has negative result of pathological histological examination.

The experiment adopts an alcoholic liver injury model method, and is provided with a low dose group, a medium dose group and a high dose group, and a blank control group and a model control group at the same time. The experimental results show that: compared with a blank control group, the average values of Malondialdehyde (MDA) Triglyceride (TG) and cell steatosis score of the mouse liver of the model control group are obviously improved, the reduced Glutathione (GSH) is obviously reduced, and the differences are obvious, which indicates that the alcoholic liver injury model is established. Compared with a model control group, the contents of MDA and TG in liver tissues of mice in the medium and high dose groups are obviously reduced, and GSH in the medium dose group is obviously increased, and the difference is obvious. The compound preparation prepared in the embodiment 1 has an auxiliary protection effect on alcoholic liver injury of mice.

The above-mentioned embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above-mentioned embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and they are intended to be included in the scope of the present invention.

TABLE 1 Effect of test substances on mouse body weight

TABLE 2 Effect of test substances on the content of MDA, GSH, TG in liver tissue of mice

Note: a. comparing the model control group with a blank control group, and carrying out t test; b. dose groupsComparing with a model control group, and carrying out variance analysis; ^# _. comparison with model control, q test, p<0.05。

TABLE 3 Effect of test substances on histopathological and histological changes of liver of mice

Note: a. comparing the model control group with a blank control group, and carrying out t test; b. dose groups were compared to model control groups, analysis of variance: 0 minute: the lipid in the liver cells is scattered and rare; l, dividing: no more than 1/4 for hepatocytes containing lipid droplets; and 2, dividing: the hepatic cells containing lipid droplets do not exceed l/2; and 3, dividing: no more than 3/4 for hepatocytes containing lipid droplets; and 4, dividing: liver tissue is almost replaced by lipid droplets.

Discussion:

at present, the prevention and treatment of alcoholic liver injury by traditional Chinese medicines are also studied, for example, patent CN202010858277.3 discloses a traditional Chinese medicine compound preparation for preventing and treating alcoholic liver injury, which is prepared by utilizing a plurality of traditional Chinese medicine materials such as herba lycopodii, herba chenopodii, cichorium intybus, rose, rhubarb, liquorice and the like. Patent CN202010787826.2 discloses a pill prepared from more than ten kinds of Chinese medicinal compositions such as radix bupleuri, radix Scutellariae, rhizoma Pinelliae Preparata, radix Glycyrrhizae Preparata and gynostemma pentaphyllum for preventing and treating alcoholic liver disease. Patent CN201910836921.4 also discloses a Chinese medicinal preparation for treating alcoholic liver disease, which is a medicament prepared from Siberian cinquefoil and its extract for treating and preventing alcoholic liver injury.

So far, the application of salvia-safflower medicine to a traditional Chinese medicine prescription as a main component for treating alcoholic liver diseases does not exist. The prescription of the compound preparation is a traditional Chinese medicine compound preparation which takes red sage roots and safflower as main materials and is assisted by vitamin C and vitamin E, wherein the compound preparation contains 350-600 parts of red sage root extract, 250-500 parts of safflower extract, 45-60 parts of vitamin C, 8-18 parts of vitamin E and a proper amount of pharmaceutic adjuvant by taking 1000 parts of the compound preparation as a basic number.

The tanshinol (derived from a salvia extract) in the formula disclosed by the invention is proved to have the liver protection effects of resisting oxidation, reducing the apoptosis and necrosis of liver cells, promoting the regeneration of the liver cells and the like. Salvianolic acid B (derived from Saviae Miltiorrhizae radix extract) has liver protecting effect on ALD rat model established by feeding alcoholic liquid feed.

Water soluble safflower yellow such as hydroxysafflower yellow A (HSYA) (derived from Carthami flos extract) can remarkably reverse the increase of ALT, AST, TG, TC, LDL in non-alcoholic fatty liver disease (NAFLD) mice, reduce liver fat content (p <0.05), and protect liver.

The vitamin C and vitamin E in the raw material components of the formulation of the invention, both of which are nutrient supplements, are one of the most important antioxidants. The combination of the salvia miltiorrhiza and safflower has the function of preventing and treating alcoholic liver injury by combining with middle polyphenol antioxidant ingredients.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes or modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the appended claims of the present application.

Claims

1. A composition comprising an active ingredient A, B, C;

2. The composition according to claim 1, wherein when said active ingredient C consists of vitamin C, vitamin E, comprising, by weight, 1000 parts per total weight of the composition:

3. A pharmaceutical formulation comprising i) the composition of claim 1 or claim 2;

ii) other pharmaceutically acceptable adjuvants, carriers or excipients.

4. Use of the composition according to claim 1 or the pharmaceutical preparation according to claim 3 for the preparation of a medicament and/or a health food for the prevention and/or treatment of alcoholic liver diseases.

5. The use according to claim 4, wherein the pharmaceutical formulation is for the manufacture of a medicament having the following effects:

(X1) reducing the level of Triglycerides (TG) in serum;

(X2) reducing hepatic Malondialdehyde (MDA) content in liver tissue;

(X3) increasing reduced Glutathione (GSH) levels in liver tissue.

6. The use of claim 4, wherein the pharmaceutical formulation is in a dosage form selected from the group consisting of: oral liquid, tablet, granule, pill, and capsule.

7. A method of preparing the pharmaceutical formulation of claim 3, comprising the steps of:

s1) providing a red sage root extract and a safflower extract;

8. The method as claimed in claim 7, wherein the step S1) of providing the salvia miltiorrhiza extract comprises the detailed steps of:

9. The method as claimed in claim 7, wherein the step S1) of providing the safflower extract comprises the detailed steps of: